Efficient generation of transgenic mice by lentivirus-mediated modification of spermatozoa

Chandrashekran, Anil, Sarkar, Rupa, Thrasher, Adrian, Fraser, Scott E., Dibb, Nicholas, Casimir, Colin M., Winston, Robert and Readhead, Carol (2014) Efficient generation of transgenic mice by lentivirus-mediated modification of spermatozoa. The FASEB Journal, 28 (2). pp. 569-576. ISSN 0892-6638

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Abstract

Transgenic technologies conventionally rely on the oocyte as a substrate for genetic modification. Owing to their accessibility, however, male germ cells, including mature sperm, have material advantages for use in transgenesis. Here we have exploited lentiviruses to generate transgenic animals via the male germline. When pseudotyped lentiviral vectors encoding green fluorescent protein (GFP) were incubated with mouse spermatozoa, these sperm were highly successful in producing transgenics. Lentivirally transduced mouse spermatozoa were used in in vitro fertilization (IVF) studies, and when followed by embryo transfer, ≥42% of founders were found to be transgenic for GFP. Inverse PCR strategy for integration site analysis demonstrated integration of at least 1 or 2 copies of GFP in the transgenics, mapping to different chromosomes. GFP expression was detected in a wide range of murine tissues, including testis and the transgene was stably transmitted to a third generation of transgenic animals. This relatively simple, yet highly efficient, technique for generating transgenic animals by transducing spermatozoa with lentiviral vectors in vitro is a powerful tool for the study of fertilization/preimplantation development, vertical viral gene transmission, gene function and regulation, and epigenetic inheritance.

Item Type: Article
Keywords (uncontrolled): lentiviral vectors, transgenesis, male germ cells, in vitro fertilization
Research Areas: A. > School of Science and Technology > Natural Sciences
A. > School of Science and Technology > Natural Sciences > Molecular Biology group
Item ID: 13936
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Depositing User: Users 3197 not found.
Date Deposited: 07 Nov 2014 17:29
Last Modified: 13 Oct 2016 14:32
URI: https://eprints.mdx.ac.uk/id/eprint/13936

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